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      Annotation of the Protein Coding Regions of the Equine Genome

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          Abstract

          Current gene annotation of the horse genome is largely derived from in silico predictions and cross-species alignments. Only a small number of genes are annotated based on equine EST and mRNA sequences. To expand the number of equine genes annotated from equine experimental evidence, we sequenced mRNA from a pool of forty-three different tissues. From these, we derived the structures of 68,594 transcripts. In addition, we identified 301,829 positions with SNPs or small indels within these transcripts relative to EquCab2. Interestingly, 780 variants extend the open reading frame of the transcript and appear to be small errors in the equine reference genome, since they are also identified as homozygous variants by genomic DNA resequencing of the reference horse. Taken together, we provide a resource of equine mRNA structures and protein coding variants that will enhance equine and cross-species transcriptional and genomic comparisons.

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          Most cited references11

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          Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.

          A new method of total RNA isolation by a single extraction with an acid guanidinium thiocyanate-phenol-chloroform mixture is described. The method provides a pure preparation of undegraded RNA in high yield and can be completed within 4 h. It is particularly useful for processing large numbers of samples and for isolation of RNA from minute quantities of cells or tissue samples.
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            The UCSC Genome Browser database: update 2011

            The University of California, Santa Cruz Genome Browser (http://genome.ucsc.edu) offers online access to a database of genomic sequence and annotation data for a wide variety of organisms. The Browser also has many tools for visualizing, comparing and analyzing both publicly available and user-generated genomic data sets, aligning sequences and uploading user data. Among the features released this year are a gene search tool and annotation track drag-reorder functionality as well as support for BAM and BigWig/BigBed file formats. New display enhancements include overlay of multiple wiggle tracks through use of transparent coloring, options for displaying transformed wiggle data, a ‘mean+whiskers’ windowing function for display of wiggle data at high zoom levels, and more color schemes for microarray data. New data highlights include seven new genome assemblies, a Neandertal genome data portal, phenotype and disease association data, a human RNA editing track, and a zebrafish Conservation track. We also describe updates to existing tracks.
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              Genome sequence, comparative analysis, and population genetics of the domestic horse.

              We report a high-quality draft sequence of the genome of the horse (Equus caballus). The genome is relatively repetitive but has little segmental duplication. Chromosomes appear to have undergone few historical rearrangements: 53% of equine chromosomes show conserved synteny to a single human chromosome. Equine chromosome 11 is shown to have an evolutionary new centromere devoid of centromeric satellite DNA, suggesting that centromeric function may arise before satellite repeat accumulation. Linkage disequilibrium, showing the influences of early domestication of large herds of female horses, is intermediate in length between dog and human, and there is long-range haplotype sharing among breeds.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                2015
                24 June 2015
                : 10
                : 6
                : e0124375
                Affiliations
                [1 ]Maxwell H. Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington, Kentucky, United States of America
                [2 ]Biochemistry and Molecular Biology Department, School of Medicine, University of Louisville, Louisville, Kentucky, United States of America
                [3 ]Department of Computer Science, University of Kentucky, Lexington, Kentucky, United States of America
                [4 ]Centre for GeoGenetics, Natural History Museum of Denmark, University of Copenhagen, Copenhagen, Denmark
                Hellas, GREECE
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: MSH JNM. Performed the experiments: SJC JNM. Analyzed the data: MSH TSK ZZ. Contributed reagents/materials/analysis tools: JL LO. Wrote the paper: MSH JNM. Critical revision of manuscript: TSK ZZ SJC JL LO.

                [¤]

                Current address: Department of Human Genetics, KU Leuven, Leuven, Belgium

                Article
                PONE-D-14-43159
                10.1371/journal.pone.0124375
                4481266
                26107351
                f94e91b8-d83c-4868-bdfb-320c6524f08a
                Copyright @ 2015

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                History
                : 29 September 2014
                : 1 March 2015
                Page count
                Figures: 4, Tables: 2, Pages: 13
                Funding
                This work was supported by the National Science Foundation (Crosscut-EF-0850237 to JL and JNM and 1054631 to JL) and a Kentucky Infrastructure for Biomedical Research Excellence award (KY-INBRE, 5P20RR016481-09) from the National Institutes of Health. Additional financial support was received from the Lourie Foundation and through endowments at the Gluck Equine Research Center, University of Kentucky. TSK’s effort was supported in part by the KY IDeA Networks of Biomedical Research Excellence (Cooper PI) NIH/NIGMS 5P20GM103436-13, and this work was conducted in part using the resources of the University of Louisville’s research computing group and the Cardinal Research Cluster. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Custom metadata
                All sequencing reads are available in the ArrayExpress database ( www.ebi.ac.uk/arrayexpress) under accession number E- MTAB-2879. A gtf format file of the transcript structures and vcf format files for transcriptomic and genomic variants are available at http://dx.doi.org/10.13013/J6057CVW.

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